Influenza A viral gene sequences are known and available and have been manipulated in the laboratory to produce influenza viruses with enhanced pathogenicity. A malicious construction and release of a highly infectious influenza virus into the population (or even the natural occurrence of a pathogenic strain) would create a pandemic that would cause the death of millions of people and seriously threaten national security. Current annual influenza vaccines are not effective from one year to the next, and thus are not effective if stockpiled. The general Aim of this proposal is to develop a stockpilable influenza vaccine for general prophylactic use comprising three plasmid DMAs (pDNAs) encoding consensus gene sequences of human influenza A nucleoprotein (NP) and Segment 7 (S7) genes and the currently circulating avian Hemagglutinin (HA) gene. The conserved NP and S7 genes would provide the generic "backbone" of the vaccine and the HA gene may require periodic updating based on the circulating pandemic strain. The completion of this project in 2 years will be accomplished by achievement of three Milestones and two Specific aims: MILESTONE 1 (to be completed prior to initiation of grant funding in July 2005): Construction of a codonoptimized HA(A/Vietnam/1203/04) pDNA and verification of its in vitro expression and mouse immunogenicity. AIM 1: Vaccination of mice and ferrets with pDNAs encoding human NP(concensus) and S7(concensus) and the avian HA(A/Vietnam/1194/04) genes and challenge the animals under BSL-3 environment with an H5N1 viral challenge. Clinical endpoint analyses will be used to evaluate antiviral efficacy. MILESTONE 2: Confirmation of anti-viral efficacy in animal(s) to be completed within 8 months of funding. AIM 2: Carry out preclinical tasks in parallel to support IND submission, including i) Manufacture of GMP pDNAs and formulate and vial vaccine; ii) mouse biodistribution study; iii) rabbit safety and toxicity dose escalation study; iv) rabbit genomic integration study; v) development of QC assays for product release; vi) development of assays to detect immunogenicity in humans. FINAL MILESTONE 3: Completion of Aim 2 and completion of IND [unreadable] [unreadable]